Hrp conjugated goat anti mouse or goat anti rabbit antibodies

Total protein was extracted by lysis for 30 min on ice in RIPA buffer pH 7.6 (150 mM NaCL, 1% Nonidet P-40, 0.5% sodium deoxycholate, 1% Triton X-100, 0.1% SDS, 1 mM EDTA, 50 mM Tris pH 7.6, 1% protease inhibitor cocktail (#P8340) and 1% phosphatase inhibitor (#P0044, both Sigma-Aldrich). Determination of protein concentrations and western blot analysis of whole-cell extracts was performed as described previously [88 (link)]. Primary antibodies were used against NRF2 (1:1000, ab62352; Abcam, Cambridge, UK), KEAP1 (sc-365626), p62/SQSTM1 (sc-28359), YAP (sc-398182; all 1:1000; Santa Cruz Biotechnology, Heidelberg, Germany), NF-κB p65 (#610868) and IKKα (#556532; both BD Bioscience, San Jose, CA, USA, 1:1000). Secondary antibodies were HRP-conjugated goat-anti-mouse or goat-anti-rabbit antibodies (1:5000, sc-2005 or sc-2004, Santa Cruz Biotechnology, Heidelberg, Germany). Anti-α-Tubulin (1:10000, B-512, Sigma-Aldrich) was used as loading control. Expression levels were visualised by WesternBright Quantum Kit (Biozym, Hessisch Oldendorf, Germany) or Super Signal West Femto (Thermo Fisher Scientific, Waltham, MA, USA) on the Li-COR C-DiGit Blot Scanner using Image Studio Digits 4.0 software (LI-COR Biosciences, Lincoln, NE, USA).